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Human CCL13/MCP-4 Antibody

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Human CCL13/MCP-4 Antibody Human CCL13/MCP-4 Antibody Monoclonal Mouse IgG1 Clone # 73506 Catalog Number: MAB327 DESCRIPTION Species Reactivity Human Specificity Detects human CCL13/MCP­4 in ELISAs. In sandwich immunoassays, less than 1% cross­reactivity with recombinant human (rh) CCL2 is observed and no significant cross­reactivity or interference with rhCCL5, 7, 8, 11, 24, recombinant mouse CCL2, 5, 7, 11 or 12 is observed. Source Monoclonal Mouse IgG1 Clone # 73506 Purification Protein A or G purified from ascites Immunogen E. coli­derived recombinant human CCL13/MCP­4 Gln24­Thr98 Accession # Q99616.1 Endotoxin Level <0.10 EU per 1 μg of the antibody by the LAL method. Formulation Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details. *Small pack size (­SP) is supplied either lyophilized or as a 0.2 μm filtered solution in PBS. APPLICATIONS Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website. Recommended Sample Concentration Intracellular Staining by Flow Cytometry 2.5 µg/106 cells A549 human lung carcinoma cell line treated with recombinant human IL­1β, fixed with paraformaldehyde and permeabilized with saponin Human CCL13/MCP­4 Sandwich Immunoassay Reagent ELISA Capture 2­8 µg/mL Human CCL13/MCP­4 Antibody (Catalog # MAB327) ELISA Detection 0.1­0.4 µg/mL Human CCL13/MCP­4 Biotinylated Antibody (Catalog # BAF327) Standard Recombinant Human CCL13/MCP­4 (Catalog # 327­P4) CyTOF­ready Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation. Neutralization Measured by its ability to neutralize CCL13/MCP­4­induced chemotaxis in the BaF3 mouse pro­B cell line transfected with human CCR2A. The Neutralization Dose (ND50) is typically 10­30 µg/mL in the presence of 2 µg/mL Recombinant Human CCL13/MCP­4. DATA Neutralization Chemotaxis Induced by CCL13/MCP­4 and Neutral­ ization by Human CCL13/ MCP­4 Antibody. Recombinant Human CCL13/ MCP­4 (Catalog # 327­P4) chemoattracts the BaF3 mouse pro­B cell line transfected with human CCR2A in a dose dependent manner (orange line). The amount of cells that migrated through to the lower chemotaxis chamber was measured by Resazurin (Catalog # AR002). Chemotaxis elicited by Recombinant Human CCL13/MCP­4 (2 µg/mL) is neutralized (green line) by increasing concentrations of Mouse Anti­Human CCL13/ MCP­4 Monoclonal Antibody (Catalog # MAB327). The ND50 is typically 10­30 µg/mL. PREPARATION AND STORAGE Reconstitution Reconstitute at 0.5 mg/mL in sterile PBS. Shipping The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (­SP) is shipped with polar packs. Upon receipt, store it immediately at ­20 to ­70 °C Stability & Storage Use a manual defrost freezer and avoid repeated freeze­thaw cycles. l 12 months from date of receipt, ­20 to ­70 °C as supplied. l 1 month, 2 to 8 °C under sterile conditions after reconstitution. l 6 months, ­20 to ­70 °C under sterile conditions after reconstitution. Rev. 2/7/2018 Page 1 of 2 Human CCL13/MCP-4 Antibody Monoclonal Mouse IgG1 Clone # 73506 Catalog Number: MAB327 BACKGROUND CCL13, also known as Monocyte Chemoattractant Protein­4 (MCP­4), is a CC chemokine that acts as a chemoattractant for monocytes, eosinophils and T cells and as an activator of basophils. Human CCL13 cDNA encodes a 98 amino acid residue precursor protein with a 23 amino acid residue hydrophobic signal peptide that is cleaved to yield an 8 kDa, 75 aa mature CCL13. Mature CCL13 lacks any potential N­glycosylation sites and shares a pyroglutamate proline motif with other human MCP proteins. Human CCL13 is most homologous to MCP­1, 3 and Eotaxin, exhibiting approximately 65­66% amino acid sequence identity. CCL13 mRNA is expressed by a number of activated cell types, including endothelial cells, macrophages, bronchial epithelium and type II alveolar cells, and perhaps lymphocytes. The bioactivities of CCL13 are mediated by the CC chemokine receptors CCR­2 and CCR­3. Rev. 2/7/2018 Page 2 of 2 .
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